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目的:初步建立蝎毒素ITX(Immunotoxin)蛋白组分的电泳分离方法和蛋白质组表达谱。方法:阴极电泳(T=18%,C=6%)对蝎毒进行分离,Bio-Rad Quantity-one电泳软件分析条带;双向电泳技术:第一向为固相pH梯度等电聚焦,第二向电泳为垂直平板SDS-PAGE(T=18%)。在以上过程中分别对上样方式、等电聚焦程序等条件进行优化控制,利用PDQ7.1软件分析获得二维凝胶图像。结果:阴极电泳图谱显示蝎毒素ITX由7个蛋白组分组成;二维凝胶图谱显示蝎毒素ITX包含相对分子质量相同(约为5.8×103)而等电点不同9种蛋白组分,其中酸性蛋白质4种,碱性蛋白质5种。结论:通过各种条件的优化,初步建立了ITX的阴极电泳和双向电泳方法,获得了分辨率和重复性较好的阴极电泳图谱和二维凝胶图谱。
OBJECTIVE: To establish an electrophoretic separation method and proteomic expression profile of protein components of ITX (Immunotoxin). METHODS: Scorpion venom was separated by cathodic electrophoresis (T = 18%, C = 6%). The band was analyzed by Bio-Rad Quantity-one electrophoresis software. The two-dimensional electrophoresis technique was as follows: Two-dimensional electrophoresis was vertical plate SDS-PAGE (T = 18%). In the above process respectively, the loading mode, isoelectric focusing procedures and other conditions for optimal control, the use of PDQ7.1 software to obtain two-dimensional gel images. Results: The results showed that ITX was composed of seven protein components. The two-dimensional gel analysis showed that ITX of scorpion toxin contained the same molecular weight (about 5.8 × 103) and different isoelectric point, of which nine protein components 4 kinds of acidic protein, 5 kinds of basic protein. Conclusion: Through the optimization of various conditions, the cathode electrophoresis and two-dimensional electrophoresis of ITX were preliminarily established, and the cathode electrophoresis pattern and two-dimensional gel map with good resolution and repeatability were obtained.